Department of Bioengineering, University of California at Berkeley, Berkeley, CA 94720, United States.
Biomaterials. 2011 Oct;32(29):6912-9. doi: 10.1016/j.biomaterials.2011.05.058. Epub 2011 Jul 20.
We have developed a synthetic polymer interface for the long-term self-renewal of human embryonic stem cells (hESCs) in defined media. We successfully cultured hESCs on hydrogel interfaces of aminopropylmethacrylamide (APMAAm) for over 20 passages in chemically-defined mTeSR™1 media and demonstrated pluripotency of multiple hESC lines with immunostaining and quantitative RT-PCR studies. Results for hESC proliferation and pluripotency markers were both qualitatively and quantitatively similar to cells cultured on Matrigel™-coated substrates. Mechanistically, it was resolved that bovine serum albumin (BSA) in the mTeSR™1 media was critical for cell adhesion on APMAAm hydrogel interfaces. This study uniquely identified a robust long-term culture surface for the self-renewal of hESCs without the use of biologic coatings (e.g., peptides, proteins, or Matrigel™) in completely chemically-defined media that employed practical culturing techniques amenable to clinical-scale cell expansion.
我们开发了一种用于在定义的培养基中长期自我更新人类胚胎干细胞（hESC）的合成聚合物界面。我们成功地在丙烯酰胺（APMAAm）水凝胶界面上培养 hESC，在化学定义的 mTeSR™1 培养基中超过 20 代，并用免疫染色和定量 RT-PCR 研究证明了多种 hESC 系的多能性。hESC 增殖和多能性标志物的结果在定性和定量上都与在 Matrigel™涂层基质上培养的细胞相似。从机制上讲，mTeSR™1 培养基中的牛血清白蛋白（BSA）对于细胞在 APMAAm 水凝胶界面上的黏附至关重要。这项研究独特地确定了一种在完全化学定义的培养基中使用实用的培养技术进行临床规模细胞扩增的无生物涂层（例如肽、蛋白质或 Matrigel™）的 hESC 自我更新的稳健长期培养表面。
